Lambda DNA mini-preps?

Anthony Palombella palomb at beagle.Colorado.EDU
Sat Dec 4 22:06:38 EST 1993


rocket1 at bu.edu (Richard Near) writes:

>If someone has a good method to prepare lambda DNA as minipreps, I'd
>be interested.  I've been growing 50ml (using LE392), doing a PEG
>precip., proteinase K, Phenol/Chloroform technique. I've been getting
>low yields, but the DNA is at least cuttable.

>  What can I do to increase my yields (I get good lysis) yet keep the 
>prep simple.  The DNA should be clean enough for clone mapping and
>I intend to use it as a template for PCR sequencing.

	Try the method of Santos (NAR 19:5442;1991).  He uses ZnCl2
to precipitate out phage particles and a simple lysis buffer to get the DNA

	Personally, I find I still need a phenol step, but that's probably
becuase I do my preps from plates (with agarose) rather than liquid more
often than not.  I usually get around 10 micrograms from a 150 mm plate,
more than enough for most diagnostic purposes.

	As long as I'm here, anyone know the chemistry involved in using
ZnCl2 to precipitate?

				-- Tony

--
```````````````````````````````````'''''''''''''''''''''''''''''''''''
HA! ...*Pronoun trouble.* It's not, "He doesn't have to shoot *you*
now," it's, "He doesn't have to shoot *me* now."  Well, I say he DOES
have to shoot me now!  So SHOOT ME NOW!! [] palomb at beagle.colorado.edu



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